Sex-specific control of human heart maturation by the progesterone receptor [bulk_RNAseq_AAV_PGR]. Sex-specific control of human heart maturation by the progesterone receptor [bulk_RNAseq_AAV_PGR]

The physiological adaptation of the heart to the postnatal environment is one of the most critical developmental transitions in the life of mammals. Despite in depth knowledge of the molecular mechanisms controlling embryonic heart development, little is known about the signals that govern postnatal maturation of the heart in humans. Here, we analyse the transcriptome of more than 50,000 single cells in the developing human heart from early gestation to adulthood, which enabled mapping of developmental trajectories across 7 main classes of cardiac cells over time. Striking sex-specific differences in cardiomyocyte maturation were identified and subsequently confirmed via deep RNA sequencing of purified cardiomyocytes. To identify transcriptional drivers of these changes, ATAC-seq was used to assay the open chromatin landscape, which unveiled the progesterone receptor as a key mediator of sex-dependent transcriptional changes during cardiomyocyte maturation. Functional studies in mice, as well as human pluripotent stem cell-derived cardiomyocytes and organoids, validated the progesterone receptor as a mediator of sex-specific metabolic programs and as a cardiac inotrope, consistent with a role in developmental maturation. These datasets provide a blueprint for understanding sex-specific mechanisms governing human heart development and unveil an important role for the progesterone receptor in cardiomyocyte maturation. Overall design: Transcriptional analysis of cardiomyocytes from mouse strain treated with 2 different adeno-associated viral vectors.

心脏对产后环境的生理适应，是哺乳动物生命周期中最为关键的发育转变之一。尽管学界对调控胚胎心脏发育的分子机制已有深入了解，但目前对于人类心脏出生后成熟过程的调控信号仍知之甚少。本研究对从妊娠早期至成年阶段的发育中人心脏内的5万余个单细胞转录组（transcriptome）进行分析，借此绘制了7大类心脏细胞随时间变化的发育轨迹。研究人员发现了心肌细胞（cardiomyocyte）成熟过程中显著的性别特异性差异，并通过对纯化心肌细胞的深度RNA测序验证了这一发现。为鉴定这些变化的转录调控因子，本研究采用转座酶可及性测序（ATAC-seq）分析染色质开放景观，揭示出孕酮受体（progesterone receptor）是心肌细胞成熟过程中性别依赖性转录变化的关键介导因子。通过小鼠实验以及人类多能干细胞（pluripotent stem cell）诱导的心肌细胞和类器官（organoids）的功能研究，验证了孕酮受体可介导性别特异性代谢程序，并作为心脏正性肌力因子发挥作用，这与其在发育成熟过程中的功能角色相符。本数据集为解析调控人类心脏发育的性别特异性机制提供了蓝图，并揭示了孕酮受体在心肌细胞成熟过程中的重要作用。总体实验设计：对经两种不同腺相关病毒载体（adeno-associated viral vector）处理的小鼠品系的心肌细胞开展转录组分析。